Attenuation of an avian pathogenic Escherichia coli strain due to a mutation in the rpsL gene

The diseases caused by pathogenic Escherichia coli constitute a major economic loss to the poultry industry. The development of a live oral E. coli vaccine to prevent or reduce diseases in poultry had been the objective of our work. Four spontaneous streptomycin-dependent (str-dependent) mutants were generated from a virulent avian strain that contains a mutation in the fur region of the chromosome. Genetic analysis of the mutants indicated that the str-dependent phenotype was due to a base change of C --> T at base 272 in the rpsL gene. The mutants were tested for attenuation using the day-old chick model. Day-old birds, in groups of 20, were either challenged with 10(6) colony-forming units (CFU) of the str-dependent mutant, the parent strain (containing the fur mutation), or the wild-type strain without the fur mutation. The parent strain and the wild-type strain were highly virulent, and 80% or more of the birds died. None of the birds challenged with the str-dependent mutants died, indicating attenuation of the mutants. The protective effect of the mutant as a live vaccine against the challenge with 10(6) CFU of the wild-type strain EC317 was investigated. Vaccination by both aerosol (day 1) and oral (days 14 and 28) routes using 10(8) CFU of the str-dependent mutant (EC1598) had no effect on the occurrence of cellulitis in the birds. Two vaccinations given as aerosol on day 1 and given orally on day 14 also had no significant effect on the occurrence of systemic lesions. Three immunizations on days 1, 14, and 28 resulted in a significant reduction in the number of birds with systemic lesions. Antibody titers prior to challenge were not predictive of outcome of challenge.     

Field-applying an inexpensive, 13C-depleted,labile carbon source to study in situ fate and short-term effects on soils

Background

Labile carbon (C_labile) limits soil microbial growth and is critical for soil functions like nitrogen (N) immobilization. Most experiments evaluating C_labile additions use laboratory incubations. We need to field-apply C_labile to fully understand its fate and effects on soils, especially at depth, but high cost and logistical difficulties hinder this approach.

Aims

Here, we evaluated the impact of adding an in situ pulse of an inexpensive and ¹³C-depleted source of C_labile—crude glycerol carbon (C_glyc), a by-product from biodiesel production—to agricultural soils under typical crop rotations in Iowa, USA.

Methods

We broadcast-applied C_glyc at three rates (0, 216, and 866 kg C ha⁻¹) in autumn after soybean harvest, tracked its fate, and measured its impact on soil C and N dynamics to four depths (0–5, 5–15, 15–30, and 30–45 cm). Nineteen days later, we measured C_glyc in microbial biomass carbon (MBC), salt-extractable organic C, and potentially mineralizable C pools. We paired these measurements with nitrate N (NO₃⁻–N) and potential net N mineralization to examine short-term effects on N cycling.

Results

C_glyc was found to at least 45-cm depth with the majority in MBC (18%–23% of total C_glyc added). The δ¹³C values of the other measured C pools were too variable to accurately track the C_labile fate. NO₃⁻–N was decreased by 13%–57% with the 216 and 866 kg C ha⁻¹ rates, respectively, and was strongly related to greater microbial uptake of C_glyc (i.e., immobilization via microbial biomass). Crude glycerol application had minor effects on soil pH—the greatest rate decreased pH 0.18 units compared to the control.

Conclusions

Overall, glycerol is an inexpensive and effective way to measure in situ, C_labile dynamics with soil depth—analogous to how mobile, dissolved organic C might behave in soils—and can be applied to rapidly immobilize NO₃⁻–N.     

Monitoring changes in back fat thickness and its effect on the restoration of ovarian activity and fertility in Bos indicus cows

With the objective of testing the hypothesis if animals with a stable layer of body fat (FAT) during the peripartum have a better chance of becoming pregnant after calving, fifty‐nine multiparous Brahman cows in their last trimester of pregnancy were used. Animals averaged four parturitions and were stocked at a rate of 1.25 animal units per hectare and divided into two groups depending on the time postpartum (dpp) that the intravaginal releasing device CIDR was inserted; Group 1 (<30 dpp; n = 30) received the implant at 25.2 ± 4.21 and withdrawn 9 days later. Group 2 (≥30 dpp; n = 29) received the CIDR at 38.41 ± 5.8. Animals were AI at detected oestrus until 170 dpp and calculated as pregnant at first service or requiring more than one service (1s and >1s), not pregnant but cycling (not pregnant) and those not cycling at all (anestrus). The FAT measurements were taken twice each month from the last trimester of gestation until 96 dpp. The onset of ovarian activity was monitored through blood levels of progesterone (P4) at days 14 and 9 prior to CIDR insertion and days 10, 13, 30 and 33 after CIDR withdrawal. Animals pregnant did not have any major changes in their fat thickness. In contrast, cows pregnant in the group ≥30 dpp had changes in their FAT homoeostasis, and pregnant animals in the 1s and >1s groups did not show differences in dorsal back fat in the last trimester of pregnancy and early postpartum. In contrast, animals not pregnant and in anestrus FAT values decreased considerably after parturition. Overall, fertility was 49%, but 18% of all the animals remained anestrus losing FAT. Thus, animals with adequate metabolic conditions will have a better chance of pregnancy regardless of the time postpartum when the reproductive programme starts.     

Estradiol and progesterone concentrations resulting from the administration of an exogenous hormone regimen in diestrous embryo transfer recipients

Estradiol and progesterone concentrations were evaluated from diestrous embryo transfer recipient mares (5 to 14 days post-ovulation) which were treated with an exogenous hormone regimen. Upon detection of the donor mare's ovulation (0 hours), 10 mg PGF_2α was given to the recipient mare; at 12, 24 and 36 hours 20 mg estradiol cypionate; at 48 hours, 500 mg progesterone in oil and then 22 mg altrenogest at 60, 72 and 96 hours. Altrenogest (22 mg/day) was continued until end of the trial (detection of a fetal heart beat). Embryos were transferred non-surgically 6 or 7 days after the start of treatment.Plasma samples were evaluated over three periods; period 1-between recipient mare ovulation and prior to PGF_2α period 2-between PGF and embryo transfer and period 3-post-transfer. During periods 2 and 3, estradiol was higher (P<.05) for mares which were 10 to 14 days post-ovulation (late diestrous) as compared to mares which were 5 to 9 days post ovulation (mid-diestrous) when treatment began. Progesterone concentrations were higher (P<.05) for the mid-diestrous mares in the same periods. The pregnancy rate was higher for the late diestrous mares than the mid-diestrous mares (58% (7/12) vs 10% (1/10)). However, no difference (P>.05) was detected in estradiol or progesterone in the late diestrous mares which were pregnant or open. During period 2, estradiol was higher (P<.05) in the pregnant than open mares. Whereas, during period 3, progesterone was higher (P<.05) in the open mares.These data suggest that estradiol is important for the establishment of pregnancy in the mare. Furthermore, hormone treatment developed in this study appears to have some potential in synchronization of diestrus mares to be used as embryo recipients.     

Spatial Response of Mammals to Late Quaternary Environmental Fluctuations

Analyses of fossil mammal faunas from 2945 localities in the United States demonstrate that the geographic ranges of individual species shifted at different times, in different directions, and at different rates in response to late Quaternary environmental fluctuations. The geographic pattern of faunal provinces was similar for the late Pleistocene and late Holocene, but differing environmental gradients resulted in dissimilar species composition for these biogeographic regions. Modern community patterns emerged only in the last few thousand years, and many late Pleistocene communities do not have modern analogs. Faunal heterogeneity was greater in the late Pleistocene.     

Rapid viral induction of plasmacytomas in pristane-primed BALB-c mice

Strain BALB/c mice were injected intraperitoneally with 0.5 milliliter of pristane, and 39 to 56 days later they were infected with Abelson murine leukemia virus, which is a lymphosarcomagenic variant of Moloney virus. Fifty-eight percent of the mice developed lymphosarcoma, and 28 percent developed immunoglobulin-producing plasmacytomas within 20 to 93 days (77 to 149 days after the pristane injection). Two of 57 control mice developed plasmacytomas at days 138 and 166 after a single injection of pristane; no plasmacytomas were found in mice treated with virus alone.     

Absorption of solar radiation by clouds: observations versus models

There has been a long history of unexplained anomalous absorption of solar radiation by clouds. Collocated satellite and surface measurements of solar radiation at five geographically diverse locations showed significant solar absorption by clouds, resulting in about 25 watts per square meter more global-mean absorption by the cloudy atmosphere than predicted by theoretical models. It has often been suggested that tropospheric aerosols could increase cloud absorption. But these aerosols are temporally and spatially heterogeneous, whereas the observed cloud absorption is remarkably invariant with respect to season and location. Although its physical cause is unknown, enhanced cloud absorption substantially alters our understanding of the atmosphere's energy budget.     

The effect of route and dosage of immunization on the serological response to a Pasteurella haemolytica and Haemophilus somnus vaccine in feedlot calves

The effect of route and dosage of administration on the serological response to a vaccine containing genetically attenuated leukotoxin of Pasteurella haemolytica combined with bacterial extracts of P. haemolytica and Haemophilus somnus (Somnu-Star Ph, Biostar Inc., Saskatoon, Saskatchewan) was evaluated in a controlled field trial in 301 feedlot calves. Vaccination of calves on arrival at the feedlot with Somnu-Star Ph significantly (p < 0.05) increased P. haemolytica and H. somnus serum antibody titers and reduced bovine respiratory disease (BRD) morbidity. A single subcutaneous vaccination with Somnu-Star Ph was as effective in stimulating a humoral antibody response and in reducing BRD morbidity as double vaccination by the intramuscular or the subcutaneous route. Furthermore, there were no swellings or adverse reactions observed with either subcutaneous or intramuscular administration of Somnu-Star Ph.

These results suggest that feedlot calves can be immunized subcutaneously once on arrival with Somnu-Star Ph. Double vaccination was of no added value in this trial, because the majority of BRD morbidity occurred prior to revaccination fourteen days postarrival. Additional larger-sized field trials are needed to monitor the duration of immunity following vaccination and to test the effect of route and dosage of vaccination on mortality.

     

Characterization of Escherichia coli isolated from cases of avian colibacillosis.

Forty-four western Canadian isolates of Escherichia coli associated with colibacillosis of turkeys and chickens were examined for serotype, antibiotic resistance, and production of aerobactin. The isolates belonged to fourteen O serogroups, with 39% of the strains being non-typeable. A high frequency of resistance to tetracycline, kanamycin, neomycin, cephalothin, streptomycin and erythromycin was observed. Most isolates produced aerobactin. Ten E. coli belonging to serogroups O1, O2 and O78 were also examined for pili production, hemagglutination, serum sensitivity, production of iron-regulated outer membrane proteins (IROMPS), and virulence. All isolates examined produced pili, exhibited mannose-sensitive hemagglutination of avian red blood cells and produced IROMPS under iron-restricted growth conditions. The five isolates of serogroup O1 and O2 were resistant to killing by turkey serum and were highly virulent. Only two of the five isolates of serogroup O78 were serum resistant. No correlation between serum resistance and virulence was observed in serogroup O78.     

Arthroscopic treatment for cervical articular process joint osteochondrosis in a Thoroughbred horse

A yearling Thoroughbred colt was presented for investigation of neck stiffness and episodes of intermittent neck pain without neurological signs. Osteochondrosis (OCD) of the cervical articular process joints (APJs) was diagnosed with the aid of radiography and computed tomography. An articular osteochondral fragment of the left fourth caudal cervical articular process was removed arthroscopically following a cut down approach to the joint capsule of the affected APJ. Surgical removal resulted in resolution of clinical signs at 4 weeks. However, subsequently the horse was markedly ataxic 6 weeks post surgery after being found cast in its stable. Cervical stenotic myelopathy was considered the most likely cause based on clinical and radiographic signs and the horse was subjected to euthanasia due to a poor prognosis for racing. Post‐mortem examination identified atypical cartilage within several cervical APJs with histological changes consistent with OCD. This case report supports OCD of the APJs as a cause of neck pain and confirms the clinical practicality and short‐term effectiveness of arthroscopic fragment removal. Surgical treatment for cervical OCD should be considered in horses without neurological signs, although case selection is important and the underlying pathology remains a risk factor for the development of subsequent neurological signs.